Cell lines that secrete anti-angiogenic antibody-scaffolds and soluble receptors and uses thereof

Abstract

The invention provides nucleic acid and polypeptide sequences encoding antibody based scaffolds such as full antibodies, antibody Fab fragments, single chain antibodies, soluble VEGF receptor-Fc fusion proteins, and/or anti-angiogenic PDGF receptors. Also encompassed are cell lines encoding such anti-angiogenic antibody scaffolds, VEGF receptors, and/or PDGF receptors. The invention also provides encapsulated cell therapy devices that are capable of delivering such anti-angiogenic antibody scaffolds, VEGF receptors, and/or PDGF receptors as well as methods of using these devices to deliver the anti-angiogenic antibody scaffolds, VEGF receptors, and/or PDGF receptors to medically treat disorders in patients, including ophthalmic, vascular, inflammatory, and cell proliferation diseases.

We claim: 
     
       1. A method for treating ophthalmic disorder associated with ocular neovascularization, comprising implanting an implantable cell culture device into the eye of a patient, wherein said device comprises;
 (a) a core comprising; 
 (i) a cell line comprising an ARPE-19 cell genetically engineered to produce a therapeutically effective amount of one or more anti-angiogenic polypeptides or anti-angiogenic molecules, wherein an iterative transfection process is used to genetically engineer the ARPE-19 cell, wherein the iterative transfection comprises one transfection, two sequential transfections, or three sequential transfections and wherein the one or more anti-angiogenic polypeptides or anti-angiogenic molecules is encoded by the nucleic acid sequence as set forth in SEQ ID NO: 1 or comprises the amino acid sequence as set forth in SEQ ID NO:2, or 
 (ii) a cell line comprising an ARPE-19 cell genetically engineered to produce a therapeutically effective amount of one or more anti-angiogenic polypeptides or anti-angiogenic molecules, wherein the therapeutically effective amount is at least 10,000 ng/day/10 6  cells, wherein the one or more anti-angiogenic polypeptides or anti-angiogenic molecules is encoded by the nucleic acid sequence as set forth in SEQ ID NO: 1 or comprises the amino acid sequence as set forth in SEQ ID NO: 2; and 
 (b) a semi-permeable membrane surrounding the ARPE-19 genetically engineered cell, wherein the membrane permits the diffusion of the one or more anti-angiogenic polypeptides or anti-angiogenic molecules there through, and 
 wherein said implanted cell culture device into the eye allows the one or more anti-angiogenic polypeptides or anti-angiogenic molecules to diffuse from the device and bind to VEGF in the eye, wherein said implantable cell culture device is implanted intraocularly or periocularly, thereby treating the ophthalmic disorder. 
 
     
     
       2. The method of  claim 1 , wherein the ophthalmic disorder is selected from the group consisting of neovascular retinopathy of prematurity, neovascular diabetic macular edema, neovascular diabetic retinopathy, neovascular age-related macular degeneration, neovascular glaucoma, neovascular retinitis pigmentosa, neovascular cataract formation, retinoblastoma neovascualrization and neovascular retinal ischemia. 
     
     
       3. The method of  claim 2 , wherein the neovascular age-related macular degeneration is wet form age-related macular degeneration. 
     
     
       4. The method of  claim 1 , wherein the ophthalmic disorder is diabetic retinopathy. 
     
     
       5. The method of  claim 1 , wherein between 0.1 pg and 1000 pg per eye per patient per day of the anti-angiogenic polypeptides or the anti-angiogenic molecule diffuses into the eye, wherein the anti-angiogenic molecule is a soluble VEGF receptor.